阳离子聚合物介导下白细胞介素1受体拮抗剂基因兔角膜原位转染及其表达

目的探讨阳离子聚合物即线性聚乙烯亚胺(PEI)介导下兔角膜基质注射PEGFP-IL- 1ra质粒进行基因角膜原位转染的有效性和安全性。方法以人cDNA文库为模板进行聚合酶链反应(PCR),获得人IL-1ra cDNA片段,构建重组质粒PEGFP-hIL-1ra。以阳离子聚合物为介导转染角膜内皮细胞,通过绿色荧光蛋白(GFP)示踪、蛋白免疫印迹技术检测转染后IL-1ra基因和蛋白的表达。实验组30只Wistar大鼠角膜基质注射PEGFP-hIL-1ra质粒和PEI-in-vivo的混合溶液20μl(含10μg质粒),对照组15只Wistar大鼠角膜基质注射PEI-in-vivo溶液20μl。注射后1、3、6、14、21 d,收集角膜通过HE染色、透射电镜、锥虫蓝-茜素红染色、免疫组织化学观察1L-1ra基因角膜原位转染后的细胞结构和功能的变化。荧光显微镜下追踪IL-1ra-GFP融合蛋白在角膜的表达部位和表达强度。结果以cDNA文库为模板扩增出hIL-1ra cDNA片段,构建重组质粒PEGFP-hIL-1ra。经PstI和BamHI酶切及DNA测序证实了插入片段方向和大小正确。PEI-in-vitro介导下PEGFP-hIL-1ra转染角膜内皮细胞12 h后,可见10％-15％细胞中有GFP荧光表达。Western-blotting检测可见相对分子质量为44 000的hIL-1ra-GFP蛋白表达。PEGFP-hIL-1ra质粒和PEI-in-vivo角膜基质注射后1 d,角膜上皮基底细胞可见荧光条带,6 d时全角膜荧光强度达到高峰,14 d开始减弱,21d角膜上皮层尚存微弱荧光。对照组观察期内始终未见绿色荧光。实验组角膜HE染色未见病理性改变,角膜上皮基底细胞层p63抗体阳性;锥虫蓝-茜素红联合染色未见角膜内皮细胞损伤;透射电镜显示角膜各层细胞的细胞质内可见IL-1ra-GFP颗粒,未见细胞器的损害。结论阳离子聚合物介导下角膜基质注射PEGFP-hIL-1ra质粒可快速、有效地将IL-1ra基因转入角膜并表达,为临床上使用抗炎细胞因子IL-1ra对角膜免疫炎性反应相关疾病进行基因治疗提供了新的技术平台

Revision of the ascomycete genus: Amphisphaerfia

published_or_final_versionEcology and BiodiversityDoctoralDoctor of Philosoph

Evaluation of Laser and Radiofrequency Induced Dorsal Root Entry Zone Lesion for Pain Control in Rats

　　Background. Dorsal root entry zone (DREZ) lesions have been believed to be effective for control of intractable pain. These lesions are usually made using radiofrequency ( RF) technique. Theoretically, laser can provide very fast, precise, reproducible and easy control of photothermal effect, possibly achieving better pain control than RF. The objective here was to learn more about the effect of pain control among the RF, carbon dioxide (CO2) laser and potassium titanyl phosphate (KPT) laser in rats. 　　Methods. The adult rat was anesthetized and the dorsal spinal cord from C5 to T1 was exposed under a microscope. The DREZ lesions were created in each group of eight including sham, RF thermocoagjlation, CO2 laser and KTP laser. The latency of pain withdrawal in the fore-paw by a hot-plate test was recorded before the DREZ lesions and three weeks afterward. 　　Result. The data showed that RF, CO2 and KTP laser could significantly reduce pain in rats ( p<0.05, one-way ANOVA and Dunnett's test). Latencies of pain withdrawal in the fore-paw by the hot-plate test before, and three weeks after , DREZ lesions were 13.9±1.4 sec and 65.2±4.9 sec in the RF group, 15.9±1.4 sec and 59.0±5.9 sec in the CO2 laser group, and 14.1±0.9 sec and 64.8±5.7 sec in the KTP group, respectively. 　　Conclusions. The density of opioid receptor in DREZ lesions cord showed no significant change three weeks after operation in the sham and CO2 laser groups. It was concluded that DREZ lesions caused by RF, CO2 laser and KTP laser can achieve pain control significantly in the rats. The effect KTP laser was close to RF, followed by CO2 laser

兰州重离子研究装置（HIRFL）感生放射性剂量率的测量

本文报道兰州重离子研究装置（ＨＩＲＦＬ）实验终端钢筒表面和实验靶处的感生放射性外照射剂量率测量的方法和结果，分析了感生放射性剂量率的衰减规律。根据实测数据评估了操作人员所受的外照射剂量，并提出了为降低操作人员受照剂量而采取的防护措施。The measurement methods and results of the dose rates from induced radioactivi-ties on the surfaces of stainless steel tubes and some experimental targets in HLRFL are pre-sented in this paper.The decay curves of induced radioactivities were analysed.Based on theexperimental data,the doses of operators were estimated,and protection methods were sug-gested in order to decrease doses to operators

黄土丘陵枣林休眠期土壤水分损失及其剖面气态水分析

针对黄土丘陵半干旱区枣林地土壤干化缺水严重的现象，利用 ２０１２ &mdash; ２０１５ 年土壤水分、温度等实测 数据，分析枣林土壤水分损失的运动规律及其机理。结果表明：休眠期是枣林地土壤水分损失十分严重的 阶段， ０ &mdash; ２００ｃｍ 土层土壤水分损失量达到 ８５．６４～９２．３４ｍｍ ，大约是同期降雨量的 ２ 倍。蒸发是休眠期 土壤水分损失的主体。在垂直剖面上，休眠期土壤水分损失自上而下呈现减少趋势，土壤水分由地下向上 运移，最终在近地表以气态水形式散失到大气。土壤气态水运动的活跃层在 ０ &mdash; １１ｃｍ 土层间，最大深度 为 ５４０ｃｍ 。受温度、相对湿度等因素的影响，土壤气态水通量随土壤深度增加而减少。</p